Multiplex polymerase chain reaction for the simultaneous detection and typing of polyomavirus JC, BK and SV40 DNA in clinical samples

Fedele, Cesare Giovanni; Ciardi, Maria; Delia, S.; Echevarría, José Manuel; Tenorio, Antonio

doi:10.1016/s0166-0934(99)00095-6

Cited by 71 publications

(46 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This histology is typical of PML. The JC virus genome was demonstrated by PCR 14 in the brain tissue. At the time of PML diagnosis, the lymphoma remained in complete remission, and the CD4 + lymphocyte count was 535/mm 3 .…”

Section: Case Reportmentioning

confidence: 99%

Progressive multifocal leukoencephalopathy after stem cell transplantation, unsuccessfully treated with cidofovir

Osorio

Cámara

Golbano

et al. 2002

Bone Marrow Transplant

View full text Add to dashboard Cite

Summary:We report a patient with progressive multifocal leukoencephalopathy (PML) after autologous stem cell transplantation (SCT) for non-Hodgkin's lymphoma (NHL). This is an unusual association, and to date only seven cases have been reported. This is the first case of PML after SCT treated with cidofovir, and the fifth case treated with this drug in a patient without human immunodeficiency virus (HIV) infection. In the previous four patients treated with cidofovir the outcome was discouraging, as was the case in this patient.

show abstract

Section: Case Reportmentioning

confidence: 99%

Progressive multifocal leukoencephalopathy after stem cell transplantation, unsuccessfully treated with cidofovir

Osorio

Cámara

Golbano

et al. 2002

Bone Marrow Transplant

View full text Add to dashboard Cite

show abstract

“…We compared the performance of the triplex qPCR assay in both formats to that of a triplex nested conventional PCR assay (15) in both single-and double-positive samples. Using the LightCycler 480 configuration, the sensitivity for JCPyV was 97.2% in single infections but 100% when both PyVs were present ( Table 4).…”

Section: Resultsmentioning

confidence: 99%

“…Results generated by using the triplex qPCR method were compared to a triplex nested conventional PCR that detects BKPyV, JCPyV, and simian virus 40 (SV40) and differentiates the amplicons by size (15). Conventional nested PCR used an ABI GeneAmp 9700 thermocycler (ABI), and second-round PCR products were visualized by electrophoresis on a 2% agarose gel stained with ethidium bromide.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Internally Controlled Triplex Quantitative PCR Assay for Human Polyomaviruses JC and BK

2008

View full text Add to dashboard Cite

We have developed a triplex TaqMan-based quantitative PCR assay for the human polyomaviruses JC (JCPyV) and BK (BKPyV). The assay simultaneously detects and quantifies both JCPyV and BKPyV in human clinical samples, and it includes an internal amplification control consisting of murine polyomavirus (MuPyV) plasmid DNA. We developed the assay for the Roche LightCycler 480 platform with the reporter dyes VIC, 6-FAM, and Cy5 for MuPyV, BKPyV, and JCPyV, respectively. The assay had a high specificity for BKPyV and JCPyV when either viral genome was present alone or in mixed samples over a range of 10 1 to 10 7 copy numbers per reaction. The analytical sensitivity was 50 copies for BKPyV and 10 copies for JCPyV. The use of the MuPyV internal control ensured monitoring of the quality of the extraction and of PCR inhibition, even in samples such as cerebrospinal fluid and plasma in which controls based on host genes cannot be effectively used. In addition, we developed a similar assay using a different dye configuration (6-FAM, VIC, and NED) that could be used on an ABI 7500 Fast platform. This assay had sensitivities similar to those of the LightCycler 480 configuration for BKPyV and JCPyV when either viral genome was present alone, but the sensitivity of detection of BKPyV was greatly decreased when an excess of JCPyV (>100-fold) was present in the sample. This internally controlled combined assay offers greater convenience and cost-effectiveness compared to separate assays for each virus and can also detect unexpected PyV activations by testing for both viruses in all samples.

show abstract

“…PCR for the DNA of BK virus and JC virus was conducted according to the method of Fedele et al 13) The products (353 bp for BKV and 189 bp for JCV) were analyzed by electrophoresis. For identification of CMV, the method of Yamamoto et al with the external primers MIE-4 and MIE-5 and the internal primers IE-1 and IE was used.…”

Section: Viral Study Of the Urine Of The Hemorrhagic Cystitis Patientsmentioning

confidence: 99%

Viruria in Adult Hemorrhagic Cystitis Patients Following Allogeneic Hematopoietic Stem Cell Transplantation and Implication of Antiviral Treatment

Kim

Lee

et al. 2007

Korean J Hematol

View full text Add to dashboard Cite

Multiplex polymerase chain reaction for the simultaneous detection and typing of polyomavirus JC, BK and SV40 DNA in clinical samples

Cited by 71 publications

References 36 publications

Progressive multifocal leukoencephalopathy after stem cell transplantation, unsuccessfully treated with cidofovir

Progressive multifocal leukoencephalopathy after stem cell transplantation, unsuccessfully treated with cidofovir

Internally Controlled Triplex Quantitative PCR Assay for Human Polyomaviruses JC and BK

Viruria in Adult Hemorrhagic Cystitis Patients Following Allogeneic Hematopoietic Stem Cell Transplantation and Implication of Antiviral Treatment

Contact Info

Product

Resources

About